83 results about "Mycobacterium fortuitum" patented technology

The invention belongs to the field of diagnostic reagents, and relates to a diagnostic reagent containing mycobacterium tuberculosis Rv1985c protein and a kit. The diagnostic reagent and the kit can perform rapid diagnosis on whether the blood or the body fluid of an experimenter is infected by the mycobacterium tuberculosis in the time periods from 15 minutes of an immune colloidal gold to 5 hours of ELISA. When the diagnostic reagent is used for the clinically-proved abortive tuberculosis, a Rv1985c antigen is used singly to detect that the sensitivity reaches 59 percent and the specificityreaches 96 percent; and when used together with other diagnostic antigens (such as a control antigen LAM / 38kDa), the diagnostic reagent can further improve the diagnostic sensitivity to 75 percent and has high clinical application values. The detection with the diagnostic reagent only needs a single blood serum sample, is simple and quick in test, needs no specialized laboratory equipment, is lowin cost, offers test results in the same day, and is very suitable for the detection of tuberculosis infection in extensive rural hospitals of villages and towns and under the condition of battlegrounds.

The invention relates to a CRISPR detection primer set for MTBC (Mycobacterium Tuberculosis Complex) and use thereof, and belongs to the technical field of gene detection. The primer set includes an amplification primer pair and a crRNA; the amplification primer pair is used to amplify a common conserved sequence of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti; the crRNA includes an anchor sequence and a guide sequence, wherein the anchor sequence is bound to a cas protein, and the guide sequence is matched with a targeted RNA fragment in the common conserved sequence.Through the primer set based on CRISPR detection technology, the detection of common conserved sequences of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti by CRISPR detection canquickly perform on-site detection of MTBC, and have the advantages of high specificity, high sensitivity and simplicity.

The invention discloses a primer and a probe for on-site rapid detection of mycobacterium tuberculosis complex and a kit thereof. The forward primer sequence is shown as SEQ ID No.1, the reverse primer sequence is shown as SEQ ID No.2, and the probe sequence is shown as SEQ ID No.3. The invention provides the PA-LFD primer and the probe for on-site distinctive, sensitive, simple and rapid detection of the mycobacterium tuberculosis complex (mycobacterium tuberculosis, mycobacterium bovis, African mycobacteria and Canna mycobacteria) and the kit containing the primer and the probe.

The present invention belongs to the field of gene engineering and diagnosis reagent technology, and aims at providing specific Rv3425 antigen for detecting tuberculosis and superior to available tuberculosis detecting reagent. The present invention provides specific Rv3425 antigen with high sensitivity in detecting pulmonary tuberculosis and other kinds of tuberculosis. The specific Rv3425 antigen as one immune dominant B-cell target antigen is used in preparing tuberculosis detecting reagent kit with high sensitivity, high safety and high reliability.

The invention discloses primers and a probe assembly for rapidly detecting mycobacterium paratuberculosis on site. A forward primer sequence is shown as SEQ ID No.1, a reverse primer sequence is shown as SEQ ID No.2, and a probe sequence is shown as SEQ ID No.3. The invention further discloses a kit for detecting the mycobacterium paratuberculosis. The primers, the probe assembly and the kit for detecting mycobacterium paratuberculosis RPA-nfo are high in sensitivity and strong in specificity, and mycobacterium paratuberculosis DNA of 8 copies / reaction can be detected at the minimum. The special instruments and equipment are not needed, by virtue of a thermostat water bath kettle or the temperature of the human armpit, the sensitive, specific and rapid detection for the mycobacterium paratuberculosis DNA can be carried out on the coarse lysate of a to-be-detected sample within 25min, and the primers, the probes and the kit are suitable for the on-site or primary-level diagnosis work of paratuberculosis.

The invention provides a method for preparing 9a-hydroxy androstendione, which comprises a step of performing fermentation culture on phytosterol by use of mycobacterium fortuitum ATCC 35855 so as to convert the phytosterol into 9a-hydroxy androstendione. The method is used for preparing 9a-hydroxy androstendione by use of mycobacterium fortuitum ATCC 35855 and corresponding fermentation liquid and fermentation technology; the cheap phytosterol can be adopted as a substrate; and moreover, the yield of the prepared 9a-hydroxy androstendione is high, a few byproducts are produced, the fermentation time is short, and a good way is provided for high-efficiency industrial production of 9a-hydroxy androstendione.

The present invention relates to diagnosis kit for Mycobacterium species identification and drug-resistance detection and manufacturing method thereof, which can discriminate a Mycobacterium Tuberculosis rpoB gene point mutation relating to the Mycobacterium species identification and drug-resistance swiftly, exactly and in large quantities using an oligonucleotide chip. The diagnosis kit for Mycobacterium species identification and drug-resistance detection in accordance with the present invention consists of an oligonucleotide chip including a Mycobacterium tuberculosis complex probe, a Mycobacterium species identification probe and a drug-resistance detection probe of a Mycobacterium tuberculosis rpoB gene, and a fluorescent material containing a biotin-binding protein so as to detect hybridization of amplified products of a specimen marked as biotine and the corresponding probe.

The invention relates to a therapeutic vaccine for malignant tumors and a composition thereof. The therapeutic vaccine for malignant tumors is a tumor cell line which contains plasmid of antisense nucleic acid of human transforming growth factor beta (TGF-beta2); the therapeutic vaccine composition for malignant tumors comprises the therapeutic vaccine for malignant tumors and an immunopotentiator, and the immunopotentiator is one selected from the group consisting of a Corynebacterium parvum preparation, a non-cell Corynebacterium parvum preparation, a BCG polysaccharide, a nucleic acid preparation, a Nocardia rubra-cell wall skeleton preparation, a group A Streptococcus preparation, a non-cell group A Streptococcus preparation, a Pseudomonas aeruginosa preparation, a non-cell Pseudomonas aeruginosa preparation, a Brucella preparation, a non-cell Brucella preparation, a non-cell Mycobacterium vaccae preparation and a non-cell Mycobacterium smegmatis preparation, preferably the non-cell Corynebacterium parvum preparation; and the malignant tumors include a lung cancer, a liver cancer, a pancreatic cancer, leukemia, lymphoma, an ovarian cancer, a colon cancer, a stomach cancer and a breast cancer.

The invention discloses a primer sequence combination and a method for applying the primer sequence combination in PCR amplification. The amplification target in the method is specific fragments EMA inside the genome of Elizabethkingia meningoseptica, and the method shows excellent specificity and sensibility; and moreover, by using the method, the neisseria meningitidis, haemophilus influenzae, staphylococcus aureus, streptococcus pneumoniae, escherichia coli, listeria monocytogenes, mycoplasma pneumoniae, bordetella pertussis, klebsiella pneumoniae and mycobacterium tuberculosis and the other pathogenic bacteria which are likely to cause meningitis and diseases of the upper respiratory tract, and easy to be confused with the Elizabethkingia meningoseptica in an identification of bacteria, can be distinguished at one time. Additionally, the lower detection limit of the method can reach 10-4 ng per Mul.

The invention belongs to the field of immunodetection and relates to a detection kit for distinguishing cow pathogenic mycobacteria infection from non-pathogenic mycobacteria infection and a method thereof. The detection reagent comprises combined fusion protein rE6-M63-H70 used as a specific stimulation origin, the combined fusion protein can effectively stimulate sensitized peripheral blood lymphocyte cultured in vitro to release Gamma-interferon (IFN-Gamma) at a high level. The cow IFN-Gamma release test established by using the detection reagent rE6-M63-H70 combined fusion protein as the stimulation origin overcomes the insufficiencies of serology detection method and the IFN-Gamma release test with PPD as the stimulation origin, thus enjoying very high sensitivity and specificity and distinguishing cow pathogenic mycobacteria ( such as mycobacterium bovis) infection from non-pathogenic mycobacteria (such as mycobacterium avium or non-pathogenic mycobacteria) infection and even distinguishing the cow pathogenic mycobacteria infection from BGG immunity; therefore, the detection kit and the method of the invention can be effectively used to detect the clinical cow pathogenic mycobacteria infection.

The invention relates to primers, probes and a method for liquid-phase chip detection of Mycobacterium tuberculosis drug-resistant gene mutation sites. The primers, probes and method are used for detecting drug-resistant gene mutation sites in Mycobacterium tuberculosis for drugs isoniazide, rifampicin and fonoquantel. The isoniazide drug-resistant mutation sites are positioned in katG gene and inhA gene; the rifampicin drug-resistant mutation sites are positioned in rpoB gene; and the fonoquantel drug-resistant mutation sites are positioned in gyrA gene. The method comprises the following steps: respectively carrying out homology analysis according to the nucleotide sequences of the four drug-resistance related genes in the gene bank, designing the primers and probes, carrying out PCR (polymerase chain reaction) twice, carrying out molecular hybridization, and carrying out detection by using a Luminex200 system, thereby determining whether the sample contains the drug-resistant mutation sites. The detection of drug-resistant gene mutation sites is of crucial importance for treating Mycobacterium tuberculosis infection by adopting correct therapeutic schedules. The primers, probes and method have the advantages of high detection speed, high sensitivity, high specificity and the like, are simple to operate, and are beneficial to popularization and application.

The invention relates to a method for extracting live bacteria RNA in Mycobacterium tuberculosis and a detection kit thereof, and particularly provides a method for extracting live bacteria RNA in Mycobacterium tuberculosis and a detection kit which is used for the live bacteria RNA in the Mycobacterium tuberculosis and is obtained by applying the method and combining fluorescent quantitative PCR technology. The kit can accurately, sensitively and quickly identify dead bacteria and the live bacteria of the Mycobacterium tuberculosis, reduce the cost and shorten the detection time, and more importantly, the kit is the basis of studies such as the diagnosis of tuberculosis, medicinal susceptibility experiments, the monitoring of chemotherapy response, the screening of new antitubercular medicaments, the prevention of the tuberculosis and the like.

The invention relates to the fields of industrial microorganisms and fermentation technology, specifically, to mycobacterium foruitum having a catalytic function and an application of mycobacterium foruitum. The mycobacterium foruitum is provided with the accession number CGMCC No.9657, and can catalyze phytosterol to produce 9[alpha]-hydroxylandrostenedione. The conversation rate is over 95%, and the selectivity is 50-60%. The unit consumption in the production of 9[alpha]-hydroxylandrostenedione through utilization of mycobacterium foruitum is 2.2-2.5, and the main product is 9[alpha]-hydroxylandrostenedione. The preparation method is simple, is high in yield, and has bright application prospects.

The invention discloses a DNA marker for detecting the drug resistance of mycobacterium tuberculosis and application of the DNA marker. It is found that mutant protein Rv2783cD67N has the DNA polymerase activity without dependence on a DNA template, is not suppressed by POA and has the phosphorolysis singe-stranded DNA (SS DNS) activity; besides, the mutant protein Rv2783cD67N can not be combined with POA; by means of the locus mutation, the drug resistance of mycobacterium tuberculosis on PZA can be remarkably caused, the locus can serve as a mark locus for molecular diagnosis of PZA drug resistance of mycobacterium tuberculosis, and the detection rate and the precision of mycobacterium tuberculosis with PZA drug resistance can be easily improved. The diagnosis time of patients can be effectively shortened, and the treatment time and cost of patients can be saved. The DNA marker for detecting the drug resistance of mycobacterium tuberculosis and application of the DNA marker provide a new thought for finding new and more effective tuberculosis drug resistance molecular markers.

The invention discloses a preparation method and application of a series of novel pyridine derivatives. The derivatives can be used for the treatment of related diseases caused by mycobacterium species, especially for diseases caused by pathogenic mycobacterium, such as Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium avium and Mycobacterium marinum.

The invention discloses a deoxyribonucleic acid (DNA) aptamer for mycobacterium tuberculosis glycolipid antigen, which has a nucleotide sequence shown as SEQ ID No. 1. Experiments show that the DNA aptamer disclosed by the invention has high specificity and strong appetency when acting with the mycobacterium tuberculosis, and a novel preparation is provided for the diagnosis of tuberculosis. Meanwhile, experiments show that the DNA aptamer can inhibit the infection of the mycobacterium tuberculosis, and a novel path is provided for the treatment of the tuberculosis.

The invention discloses a method for producing A ring degradation products through plant sterol fermentation. The method comprises the following steps: (A) preparing a liquid inoculant containing mycobacterium fortuitum; (B) carrying out fermentation culture; (C) extracting and separating so as to obtain the A ring degradation products. The provided fermentation method for preparing A ring degradation products is carried out in an oil-free condition, thus the ring opening of A ring and B ring is effectively inhibited, thus the degradation is inhibited, the yield is increased, and the weight yield can reach 35 to 40%.

The invention provides a kit for detecting an alpaca mycobacterium paratuberculosis antibody and a detection method thereof, belonging to the technical field of biology. The kit comprises mycobacterium paratuberculosis recombinant hed protein, positive control serum and negative control serum of alpaca paratuberculosis, an ELISA plate, coating liquid, confining liquid, PBST, staphylococcal protein A labeled by horseradish peroxidase (HRP-SPA), a raw material for preparing developing solution and stop solution. The kit of the invention introduces the high-expression mycobacterium paratuberculosis hed protein with strong specificity as a detection antigen for directly detecting the alpaca mycobacterium paratuberculosis antibody, thus having good specificity and sensitivity. The method for detecting the alpaca mycobacterium paratuberculosis antibody by the kit is simple, convenient, fast and accurate, thus being applicable to sample detection in batches.

The invention provides a primer combination and an application thereof. The primer combination comprises a first primer group and a second primer group, wherein the nucleotide sequence of the first primer group is shown in the SEQ ID NO:1-2, and the nucleotide sequence of the second primer group is shown in the SEQ ID NO:3-4. The primer combination can specifically recognize the mycobacteria specific area-ITS area, has high homology between different mycobacteria, and is applicable to various mycobacteria and capable of being effectively used for recognizing the mycobacteria, particularly tuberculous mycobacteria and non-tuberculous mycobacteria.

The invention provides a drug target point based on mycobacterium tuberculosis ubiquitin ligase PafA and an application thereof, and belongs to the technical field of biomedicine. Specifically, the invention relates to a use of a 119-site serine of ubiquitin ligase PafA as a target point of an anti-tuberculosis drug. The serine site at the 119 site of the protein can specifically bind to a small molecule inhibitor 4-(2-aminoethyl)benzenesulfonylfluoride hydrochloride so that the activity of the mycobacterium tuberculosis ubiquitin ligase PafA is inhibited.

The invention relates to a primer for detecting mycobacteria. Nucleotide sequences of an upstream primer and a downstream primer are shown as sequence tables, namely SEQ ID No.1 and SEQ ID No.2 respectively. A polymerase chain reaction-denatured high performance liquid chromatography (PCR-DHPLC) kit for detecting the mycobacteria comprises a PCR buffer solution, a primer pair for detecting the mycobacteria, deoxy-ribonucleoside triphosphate (dNTP), MgCl2, thermus aquaticus deoxyribonucleic acid (Taq DNA) polymerase, double distilled water, negative control (sterile physiological saline) and positive control (plasma DNA with an amplification product), wherein a nucleotide sequence of the amplification product is shown as SEQ ID No.3. The invention provides a kit for rapidly detecting the infection of the mycobacteria and a PCR-DHPLC detection method applying the kit. The kit and the detection method have high specificity, sensitivity and throughput, are easy to operate, and have a great practical significance for clinically distinguishing the infection of the mycobacteria and other pathogens.

The invention discloses a probe and a kit for the common pathogens detection of skin infectious granuloma. The probe comprises a mycobacterium probe, a mycobacterium tuberculosis probe, a mycobacterium leprae probe, a mycobacterium avium probe, a mycobacterium intracellulare probe, a mycobacterium kansasii probe and a mycobacterium fortuitum probe, a candida albicans probe, a sporotrichum schenckii probe, a cladosporium trichoides probe, a Fonsecaea pedrosoi probe, and a Fonsecaea monophora probe; and a nocardia probe, an actinomycetes probe and a leishmania probe. The nucleotide sequence of each probe is shown in sequence tables SEQ ID NO.1-SEQ ID NO.15; and the kit comprises the probes and corresponding universal primers. According to the invention, through designing and optimizing the kit for carrying out pathogens screening and detection on common skin infectious granuloma, the improvement of the clinical diagnosis and the treatment efficiency is facilitated.

The invention discloses a substituted nitroimidazole derivative, which is mainly used for treating related diseases caused by mycobacterium infection, such as Mycobacterium tuberculosis, especially for diseases caused by drug-resistant Mycobacterium tuberculosis.

The invention discloses a method for identifying mycobacterium. The method comprises the following steps: acquiring a plurality of standard sequences of a plurality of mycobacteria so as to generate a mycobacterium sequence database; by comparing the standard sequences, obtaining a conservation section mutually owned by the standard sequences; preparing a PCR primer according to the conservation section; using the PCR primer to perform the PCR amplification of a DNA solution of the testing mycobacterium so as to obtain an amplification product of the testing mycobacterium; carrying out the sequencing reaction of the amplification product to obtain the sequencing product; carrying out the sequencing analysis of the sequencing product to obtain a sequence of the testing mycobacterium; and by comparing the testing sequence with the standard sequence in the mycobacterium sequence database, determining the type of the testing mycobacterium. The PCP primer is designed according to the conservation section of the standard sequences of various mycobacteria, only a pair of primers is needed for all PCRs and sequencing, thereby greatly reducing the quantity of oligonucleotide needed for experimental population.

The invention discloses a mycobacterium tuberculosis detecting kit and an application thereof. The mycobacterium tuberculosis detecting kit comprises a coating solution, a cleaning solution, a blocking solution, a stopping solution, an antigen Rv1040c and an antigen Rv2309A. The kit disclosed by the invention has the advantages of simplicity and convenience in operation and equipment, low cost, high precision, environment friendliness and the like; according to the mycobacterium tuberculosis detecting kit, the mixed antigens are used for coating, so that the problem of large individual difference caused when a single antigen is used for diagnosis is solved; in addition, the detecting effects of the mixed antigens are superior to the detecting effect of the single antigen on the aspects of sensitivity, precision and specificity. Compared with the similar kit sold on the market at present, the mycobacterium tuberculosis detecting kit disclosed by the invention has more remarkable advantages on the aspects of specificity, false positive rate or detection rate and has a wide application prospect in detecting mycobacterium tuberculosis.

The present invention discloses a PCR-RLB assay process of mycobacteria and the employed primers and probes. A pair of oligonucleotide primers and 35 species or type-specific probes are designed against the characteristics of DNA sequences of different types of mycobacterias. Via PCR-RLB assay process, 17 mycobacterias and Mycobacterium tuberculosis complexes, Mycobacterium Ulcerans / Marinum and 2 subspecies of Mycobacterium Kansasii can be identified for SGM. 14 species or subspecies can be identified for RGM. Compared with traditional identification process of mycobacteria, the present invention is a rapid, simple, practical, specific and sensitive assay method.

The invention relates to the field of biological detection of molecules, in particular to the field of identification of mycobacteria, and provides an oligonucleotide combination. The oligonucleotidecombination comprises oligonucleotide for detecting mycobacterium fortuitum and oligonucleotide for detecting mycobacterium intracellulare, and selectively comprises at least one group, which can specifically detect mycobacterium tuberculosis and non-tuberculous mycobacteria and can identify specific species / subspecies, of the rest three groups, and meanwhile, two kinds of mycobacteria can be detected through a channel, so that the number of mycobacteria detected by fluorescence PCR is increased. The invention further provides a kit containing the oligonucleotide combination and a method usedfor identifying the mycobacteria.

The invention belongs to the technical field of steroid biotransformation, and particularly relates to a mycobacterium fortuitum capable of effectively degrading phytosterol side chains and producing9alpha-OH-AD and application thereof. The mycobacterium fortuitum is a mutagenic strain obtained by physico-chemical combination mutagenesis, and the yield of 9alpha-OH-AD is up to 97%; when the concentration of a phytosterol substrate is up to 95.7% and is 1.21 times of that of original strains, and the biotransformation time is 72 hours and is 24 hours shorter than the original strains. Comparedwith the prior art, mycobacterium fortuitum using phytosterol as the substrate for producing 9alpha-OH-AD is highest in conversion rate and shortest in conversion time, and the mycobacterium fortuitum has a good industrial application prospect.

The invention belongs to the field of biomedicines, and relates to an application of vitamin C (ascorbic acid), as a single component, in preparation of medicines for treating and preventing tuberculosis. Through cytobiology and pharmacology experiments with combination of animal experiments, anti-tuberculosis pharmaceutical concentration of the vitamin C is determined; and on the basis of regulatory mechanism on oxidative stress status of mycobacterium tuberculosis after infection by macrophages, interference test is carried out, wherein a result proves that high-dose vitamin C can kill the mycobacterium tuberculosis, so that inhibition effect is achieved through NAC and catalase; the vitamin C treats the macrophages to generate H2O2, wherein functional effect is achieved with the signalchannel being same as H2O2; large-dose vitamin C kills mycobacteria, wherein inhibition function is achieved through a glutathione precursor NAC and CAT; high-dose vitamin C can infect the TNF-alpha signal pathway induced by RAW 264.7 cells via mediation of Bacillus Calmette-Guerin vaccine and H37Rv, thus achieving sterilization effect and influence on body immunity. The invention provides effective treatment strategy for clinical therapy and drug resistance of tuberculosis and tuberculosis in incubation period.

Disclosed is a microorganism having an excellent ability to convert sterol into and rost-4-ene-3,17-dione (AD) and androsta-1,4-diene-3,17-dione (ADD), a method for preparation of the microorganism and use thereof, and more particularly, a mutant strain of Mycobacterium fortuitum ATCC 29472, Mycobacterium fortuitum EUG-119, a method for preparation of the mutant and use thereof in preparing AD and ADD. The mutant of the present invention has an excellent ability to convent sterol into AD and ADD which are steroid hormone precursors, compared with the known microorganisms and accordingly, is very useful for mass production of steroid hormones.